Safety
Introduction – German Safety Regulations
In Germany, working with genetically modified organisms is regulated by the genetic engineering law (Deutsches Gentechnikgesetz, GenTG). The primary purpose of these laws and regulations is the protection of people and the environment from potential threats.
Biosafety vs. Biosecurity
"Biosafety protects people from germs – biosecurity protects germs from people" [chabsa]
Biosafety
When discussing safe work on laboratories, it is important to understand the difference between biosafety and biosecurity. Biosafety generally refers to the protection of people inside and outside of the lab, animals and the environment. The rules of conduct for biosafety are derived from the World Health Organization (WHO) on techniques for use in laboratories. According to the WHO biosafety is “the containment principles, technologies and practices that are implemented to prevent unintentional exposure to pathogens and toxins, or their accidental release” [chabsa]. According to this principle, biosafety is about taking precautions to avoid harm from happening. While working on our project, we were highly determined never to pose any threats to ourselves or the public. All experiments were strictly contained within the lab and we adhered to laboratory code of conducts at all times. Furthermore, all experiments were thoroughly planned and modeled before conducted in the lab in order to anticipate problems. Transportation of strains etc. between labs was always performed in S1 approved safety boxes.
Biosecurity
The term biosecurity refers to the protection of biological constructs and inventions from misuse. According to the World Health Organization laboratory biosecurity describes “the protections, control and accountability for valuable biological materials within laboratories, in order to prevent their unauthorized access, loss, theft, misuse or diversion or intentional release” [chabsa].
Work Environment – Lab Safety
In order to be able to work safely in the lab and to adhere to all rules and regulations, every member received extensive safety inductions before commencing their work. First of all, we received an introduction to the various safety levels laboratories can be classified into. There are four internationally recognized safety levels:
Safety level one (S1) represents the lowest safety level and refers to work with agents that do not pose a considerable threat to people working in the lab or the environment [2]. Since there is no high risk of infections or disease, research can be performed on standard open benches and no use of special containment equipment is required. Furthermore, laboratories that are classified S1 are not isolated from the general building. All work we conducted during the iGEM competition could be done in an S1 laboratory.
Naturally, our safety induction also included general codes of conduct and standard microbiology practices, which are indispensable in order to create a safe work environment [labmanager]. These rules include:
- no mouth pipetting, mechanical pipetting only
- careful and safe handling of sharp equipment
- immediate decontamination of spills
- immediate decontamination of all surfaces when work is finished
- decontamination of waste by autoclaving
- posting of “Biohazard” signs whenever infectious substances are dealt with
Finally, in order to ensure complete protection, one has to pay attention to personal hygiene. Hand washing is required after each completed job and before leaving the lab. Furthermore, eating, drinking, smoking and applying makeup or equivalent cosmetic products are prohibited.
Personal Protection Equipment
While carrying out our work in the lab, personal protection equipment (PPE) had to be worn at all times. Since all of our work was carried out in a S1 classified lab, our PPE consisted of eye protection, a lab coat consisting of 100 % cotton, sturdy footwear and gloves. Furthermore, jewelry was taken off before entering the lab and long hair was tied back or tucked into the lab coat.
Personal safety training
We received the above mentioned safety inductions in every laboratory we worked in. They were carried out by safety advisors, Postdocs who received special safety induction training, or the lab managers themselves.
- Institute for Plant and Cell Biotechnology: Prof. Dr. Markus Pauly (Head of Institute), Fabian Stritt (PhD student)
- Institute for Microbiology: Prof. Dr. Michael Feldbrügge (Head of Institute)
- Institute for Synthetic Biology: Prof. Dr. Matias Zurbriggen (Head of Institute)
Disposal of Biological Waste
Following the safety inductions, we applied everything we learned to safely dispose of our waste. All lab ware we used (culture dishes, petri dishes, centrifuge tools, pipettes and tips, etc.) was non-infectious so that we could dispose of them in special plastic bags within the lab. These bags were autoclaved by a technical assistant of the lab. Liquid waste was collected in special containers that were later autoclaved and disposed by a technical assistant. Sharps waste, like razor blades were disposed in special sharps waste containers to prevent injuries.
Organisms and parts used in our project
Organisms
| Species Name | Strain | Genotype | Risk group | Disease risk | Source |
|---|---|---|---|---|---|
| E.coli | K-12 TOPTEN | F- mcrA Δ(mrr-hsdRMSmcrBC)φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(araleu)7697 galU galK rpsL endA1 nupG P3: Kan R; AmpR; (am) Tet; R (am) | 1 | no | Grant et al. 1990 |
| S.cerevisiae | BY4742 | MATα his3Δ1 leu2Δ0 lys2Δ0 ura3Δ0 | 1 | no | Brachmann et al. 1998 |
Parts
| BioBrick | Name | Plasmid |
|---|---|---|
| BBa_K2271066 | mRuby-ePTS1 | pSB1C3 |
| BBa_K2271062 | Pex13-mTurqoise | pSB1C3 |
| BBa_K2271060 | Snc1 | pSB1C3 |
| BBa_K2271103 | Pex15 | pSB1C3 |
| BBa_K2271104 | Pex13 membrane anchor | pSB1C3 |
| BBa_K2271105 | Pex5 variant R15 | pSB1C3 |
| BBa_K2271106 | PTS1 variant P3 | pSB1C3 |
| BBa_K2271107 | Pex5 variant 19 comp | pSB1C3 |
| BBa_K2271067 | Pex5 variant R19 | pSB1C3 |
| BBa_K2271014 | VioA | pSB1C3 |
| BBa_K2271017 | VioE | pSB1C3 |
| BBa_K2271115 | ADH alcohol dehydrogenase | pSB1C3 |
| BBa_K2271116 | ADH PTS1 | pSB1C3 |
| BBa_K2271118 | ValS | pSB1C3 |
| BBa_K2271124 | ValS PTS1 | pSB1C3 |
| BBa_K2271021 | pH sensitive green flourescent protein for yeast expression | pSB1C3 |
| BBa_K2271023 | reduction-oxidation sensitive green fluorescent protein for yeast expression | pSB1C3 |
| BBa_K2271024 | reduction-oxidation sensitive green flourescent protein with pts1 for Yeast expression | pSB1C3 |
| BBa_K2271026 | roGFP2 with pts1 and Pex13-mRuby fusion construct | pSB1C3 |
| PTS1 variant P* | pSB1C3 | |
| BBa_K2271141 | PEX13 mRuby comp | pSB1C3 |
| BBa_K2271142 | ePTS | pSB1C3 |
| BBa_K2271143 | Pex3 | pSB1C3 |
| BBa_K2271144 | Pex26 | pSB1C3 |
| BBa_K2271145 | Bacteriorhodopsin | pSB1C3 |
| BBa_K2271013 | Pex34p with Copper induecable promotor | pSB1C3 |
| BBa_K2271010 | Pex34p with ScCCW12 promoter | pSB1C3 |
| BBa_K2271008 | Pex34-mTurquoise with Galactose inducible promoter | pSB1C3 |
| BBa_K2271007 | Pex34p-mTurquoise with ScPAB1 promoter | pSB1C3 |
| BBa_K2271006 | Pex34p-mTurquoise with ScCCW12 promoter | pSB1C3 |
| BBa_K2271003 | Pex11-mVenus with galactose-inducible promoter | pSB1C3 |
| BBa_K2271002 | Pex11-mVenus with ScPAB1 promoter | pSB1C3 |
| BBa_K2271000 | Pex11 | pSB1C3 |
| BBa_K2271005 | Pex34 coding sequence | pSB1C3 |
| BBa_K2271011 | Pex34 with ScPAB1 promoter no tag | pSB1C3 |
| BBa_K2271012 | Pex34 with galactose inducible promoter | pSB1C3 |
